Fig. 4

Lymphocyte infiltration into brain, IFN-γ production and host-defense factors. Immune cells were isolated from brain, and identified as single, live, CD45^hiCD11b⁻CD3⁺ lymphocytes as shown above. A Representative gating illustrates discrimination of CD4⁺ and CD8⁺ T cells, and values in quadrant gates indicate cell frequency from parent population. B Total cell number of recruited CD4⁺ and CD8⁺ lymphocytes to the brain. C–D Intracellular analysis of IFN-γ production. Bar charts present MFI values as mean + SEM, n = 4, **p < 0.01 (unpaired two-tailed t-test). Histograms show the representative IFN-γ production by CD4⁺ and CD8⁺ T cells in comparison to isotype-control. Bars and numbers above histograms indicate mean percentage of positively stained cells + SEM. E Gene expression levels of host-defense factors in the whole brain homogenate were assessed using RT-qPCR. Bar charts represent mean values + SEM obtained in two independent experiments and analyzed together, n = 4–5 per experiment, *p < 0.05 (Mann–Whitney U-test). Control (black bars) and PACAP-treated (white bars)